[Zoonoses with wildlife reservoirs: a threat to public health and the economy].
The world is experiencing an increase in emergent infections as a result of anthropogenic changes of the biosphere and globalization. Global warming unrestricted exploitation of natural resources such as forests and fisheries, urbanization, human migration, and industrialization of animal husbandry cause environmental destruction and fragmentation. These changes of the biosphere favor local emergence of zoonoses from their natural biotopes and their interaction with domestic animals and human populations. Subsequently, international commerce, human and animal migration and travel, favor the dissemination of these zoonotic pathogens worldwide. Chile is undergoing an important degradation of many wild-life biotopes, affecting their diversity and contributing to the dissemination of zoonoses such as Chagas disease, Hantavirus, rabies, fish tapeworms, and marine vibriosis. Moreover, agents of many other zoonoses such as inverted exclamation markeptospirosis, hydatidosis, salmonellosis, rabies, brucellosis and anthrax have been detected in different wild-life environments in the country. The intensification and accelerations of the anthropogenic deterioration of the biosphere in Chile, as results of the unrestricted utilization of natural resources and global climate change, suggests that emergence of new zoonoses in the near future will lead to important public health and economic problems. Forestalling of these problems will require active epidemiológica! surveillance of wild and domestic animals with a wide range of modern molecular and ancillary epidemiológica! tools. This also demands government and private sector (i.e., animal husbandry) intervention, funding and the collaboration of professionals in human and veterinary medicine with those in the environmental sciences including ecology, climatology and oceanography.


Editor's Choice: Contemporary treatment of popliteal artery aneurysm in eight countries: A Report from the Vascunet collaboration of registries.
To study contemporary popliteal artery aneurysm (PA) repair. Vascunet is a collaboration of population-based registries in 10 countries: eight had data on PA repair (Australia, Finland, Hungary, Iceland, New Zealand, Norway, Sweden, and Switzerland). From January 2009 until June 2012, 1,471 PA repairs were registered. There were 9.59 operations per million person years, varying from 3.4 in Hungary to 17.6 in Sweden. Median age was 70 years, ranging from 66 years in Switzerland and Iceland to 74 years in Australia and New Zealand; 95.6% were men and 44% were active smokers. Elective surgery dominated, comprising 72% of all cases, but only 26.2% in Hungary and 39.7% in Finland, (p < .0001). The proportion of endovascular PA repair was 22.2%, varying from 34.7% in Australia, to zero in Switzerland, Finland, and Iceland (p < .0001). Endovascular repair was performed in 12.2% of patients with acute thrombosis and 24.1% of elective cases (p < .0001). A vein graft was used in 87.2% of open repairs, a synthetic or composite graft in 12.7%. Follow-up was until discharge or 30 days. Amputation rate was 2.0% overall: 6.5% after acute thrombosis, 1.0% after endovascular, 1.8% after open repair, and 26.3% after hybrid repair (p < .0001). Mortality was 0.7% overall: 0.1% after elective repair, 1.6% after acute thrombosis, and 11.1% after rupture. Great variability between countries in incidence of operations, indications for surgery, and choice of surgical technique was found, possibly a result of surgical tradition rather than differences in case mix. Comparative studies with longer follow-up data are warranted.


Inheritance, gene expression, and lignin characterization in a mutant pine deficient in cinnamyl alcohol dehydrogenase.
We have discovered a mutant loblolly pine (Pinus taeda L.) in which expression of the gene encoding cinnamyl alcohol dehydrogenase (CAD; EC 220.127.116.11) is severely reduced. The products of CAD, cinnamyl alcohols, are the precursors of lignin, a major cell wall polymer of plant vascular tissues. Lignin composition in this mutant shows dramatic modifications, including increased incorporation of the substrate of CAD (coniferaldehyde), indicating that CAD may modulate lignin composition in pine. The recessive cad-n1 allele, which causes this phenotype, was discovered in a tree heterozygous for this mutant allele. It is inherited as a simple Mendelian locus that maps to the same genomic region as the cad locus. In mutant plants, CAD activity and abundance of cad RNA transcript are low, and free CAD substrate accumulates to a high level. The wood of the mutant is brown, whereas the wood in wild types is nearly white. The wood phenotype resembles that of brown midrib (bm) mutants and some transgenic plants in which xylem is red-brown due to a reduction in CAD activity. However, unlike transgenics with reduced CAD, the pine mutant has decreased lignin content. Wood in which the composition of lignin varies beyond previous expectations still provides vascular function and mechanical support.


Targeted estimation of nuisance parameters to obtain valid statistical inference.
In order to obtain concrete results, we focus on estimation of the treatment specific mean, controlling for all measured baseline covariates, based on observing independent and identically distributed copies of a random variable consisting of baseline covariates, a subsequently assigned binary treatment, and a final outcome. The statistical model only assumes possible restrictions on the conditional distribution of treatment, given the covariates, the so-called propensity score. Estimators of the treatment specific mean involve estimation of the propensity score and/or estimation of the conditional mean of the outcome, given the treatment and covariates. In order to make these estimators asymptotically unbiased at any data distribution in the statistical model, it is essential to use data-adaptive estimators of these nuisance parameters such as ensemble learning, and specifically super-learning. Because such estimators involve optimal trade-off of bias and variance w.r.t. the infinite dimensional nuisance parameter itself, they result in a sub-optimal bias/variance trade-off for the resulting real-valued estimator of the estimand. We demonstrate that additional targeting of the estimators of these nuisance parameters guarantees that this bias for the estimand is second order and thereby allows us to prove theorems that establish asymptotic linearity of the estimator of the treatment specific mean under regularity conditions. These insights result in novel targeted minimum loss-based estimators (TMLEs) that use ensemble learning with additional targeted bias reduction to construct estimators of the nuisance parameters. In particular, we construct collaborative TMLEs (C-TMLEs) with known influence curve allowing for statistical inference, even though these C-TMLEs involve variable selection for the propensity score based on a criterion that measures how effective the resulting fit of the propensity score is in removing bias for the estimand. As a particular special case, we also demonstrate the required targeting of the propensity score for the inverse probability of treatment weighted estimator using super-learning to fit the propensity score.


Screening family planning needs: an operations research project in Guatemala.
BACKGROUND: Public sector health care providers in rural Guatemala have infrequently offered family planning information and services in routine visits. This operations research project tested a strategy to modify certain practices that prevent health workers from proactively screening clients' needs and meeting them. METHODS: The research design was quasi-experimental with a pretest-posttest-follow-up comparison group design. Health districts, which comprise health centers and posts, were purposively assigned to intervention or comparison groups to assure comparability of the two groups. The strategy was based on a job-aid designed to guide health workers in screening clients' reproductive intentions and family planning needs, help them to offer contraceptive methods if the woman expressed interest, and facilitate the provision of the method chosen at the time of the visit. The strategy was implemented at intervention sites during a period of six months. Upon completion of post-intervention measurements, the strategy was scaled up to the comparison sites, and a follow-up assessment was conducted nine months later. Results were evaluated by conducting three rounds of exit interviews with women exposed to the risk of unwanted pregnancy. RESULTS: Study results showed a two to five-fold increase in providers' screening of clients' reproductive intentions. The proportion of clients who received information about contraceptives increased from 8% at the baseline to 42% immediately post-intervention, and 36% at the follow-up survey. The intervention also proved successful in improving the role service providers play in offering women a chance to ask questions and assisting women in making a selection. The proportion of women who received a method, referral or appointment increased and remained high in the intervention group, although no change was seen in the comparison group after their participation in the strategy. CONCLUSION: The easy-to-use job aid developed for this project proved useful for screening clients' needs and reducing providers' reluctance to discuss family planning with clients and offer contraceptive services. Such family planning screening devices can be useful in traditional settings where both providers and clients shy away from discussing family planning issues.


Comparative genomics of serotype Asia 1 foot-and-mouth disease virus isolates from India sampled over the last two decades.
This study deals with a comparative analysis of complete genome sequences of twenty-one serotype Asia 1 foot-and-mouth disease (FMD) field viruses isolated over a period of two decades from India, two vaccine strains and seven exotic sequences. The Indian viruses could be grouped in to three distinct lineages at the entire coding region, evolving independently probably under differential selection pressure as evident from the lineage-specific signatures identified. This comparison revealed 80% of amino acids at the polyprotein region to be invariant. Twenty-one residues in L, 3A and P1 region were identified to be under positive selection of which some are antigenically critical. Analysis at functionally crucial motifs, receptor contact residues, polyprotein cleavage sites and at putative T-cell epitopes expands the knowledge beyond other serotypes. Antigenic site II in betaB-betaC loop of VP2 was highly unstable suggesting its exposure to extreme immune pressure. A single cross-over at the L proteinase region in an isolate from buffalo, also featuring an extensive deletion at the 5' untranslated region (UTR), reflects the role of intraserotypic genetic recombination in natural evolution. The likely biological relevance of deletions/insertions observed at UTRs, VP1 and 3A could not be deduced. Altogether, a substantial amount of data raised on full length genomes of type Asia 1 virus adds value to the FMD virus genomics.


Treatment of asymptomatic pulmonary cryptococcosis in immunocompetent hosts with oral fluconazole.
Pulmonary cryptococcosis is occasionally detected on routine imaging studies in healthy hosts with no or mild symptoms. Isolated pulmonary cryptococcosis may be observed without specific therapy in asymptomatic immunocompetent hosts. However, considering that dissemination from a pulmonary infection can occur in patients with no immunologic defects, treatment of asymptomatic pulmonary cryptococcosis in immunocompetent hosts remains controversial. The aim of this study was to determine the role of fluconazole therapy in the management of isolated pulmonary cryptococcosis in asymptomatic healthy hosts. We retrospectively analyzed the medical records and radiographic findings of 10 healthy subjects with isolated pulmonary cryptococcosis diagnosed incidentally and treated with oral fluconazole. All patients had no respiratory or constitutional symptoms. The most common radiological findings were pulmonary nodules, and the number of nodules in each patient was from 1 to 9. After histological confirmation, all patients were treated with oral fluconazole at a dosage of 400 mg per day for a median period of 6.4 months. No patient developed an adverse reaction to fluconazole. The mean interval between the initiation of antifungal therapy and final radiological response was 8.3 months. Seven of the 10 patients showed complete resolution, and the other 3 patients were assessed as having partial resolution. During the average follow-up period of 11.9 months, all patients showed a favourable outcome with no relapse. The overall cure rate was 70%. These results suggest that fluconazole may be an attractive therapeutic option for asymptomatic pulmonary cryptococcosis in immunocompetent hosts.


Independent association of extent of resection with survival in patients with malignant brain astrocytoma.
With recent advances in the adjuvant treatment of malignant brain astrocytomas, it is increasingly debated whether extent of resection affects survival. In this study, the authors investigate this issue after primary and revision resection of these lesions. The authors retrospectively reviewed the cases of 1215 patients who underwent surgery for malignant brain astrocytomas (World Health Organization [WHO] Grade III or IV) at a single institution from 1996 to 2006. Patients with deep-seated or unresectable lesions were excluded. Based on MR imaging results obtained < 48 hours after surgery, gross-total resection (GTR) was defined as no residual enhancement, near-total resection (NTR) as having thin rim enhancement of the resection cavity only, and subtotal resection (STR) as having residual nodular enhancement. The independent association of extent of resection and subsequent survival was assessed via a multivariate proportional hazards regression analysis. Magnetic resonance imaging studies were available for review in 949 cases. The mean age and mean Karnofsky Performance Scale (KPS) score at time of surgery were 51 +/- 16 years and 80 +/- 10, respectively. Surgery consisted of primary resection in 549 patients (58%) and revision resection for tumor recurrence in 400 patients (42%). The lesion was WHO Grade IV in 700 patients (74%) and Grade III in 249 (26%); there were 167 astrocytomas and 82 mixed oligoastrocytoma. Among patients who underwent resection, GTR, NTR, and STR were achieved in 330 (35%), 388 (41%), and 231 cases (24%), respectively. Adjusting for factors associated with survival (for example, age, KPS score, Gliadel and/or temozolomide use, and subsequent resection), GTR versus NTR (p < 0.05) and NTR versus STR (p < 0.05) were independently associated with improved survival after both primary and revision resection of glioblastoma multiforme (GBM). For primary GBM resection, the median survival after GTR, NTR, and STR was 13, 11, and 8 months, respectively. After revision resection, the median survival after GTR, NTR, and STR was 11, 9, and 5 months, respectively. Adjusting for factors associated with survival for WHO Grade III astrocytoma (age, KPS score, and revision resection), GTR versus STR (p < 0.05) was associated with improved survival. Gross-total resection versus NTR was not associated with an independent survival benefit in patients with WHO Grade III astrocytomas. The median survival after primary resection of WHO Grade III (mixed oligoastrocytomas excluded) for GTR, NTR, and STR was 58, 46, and 34 months, respectively. In the authors' experience with both primary and secondary resection of malignant brain astrocytomas, increasing extent of resection was associated with improved survival independent of age, degree of disability, WHO grade, or subsequent treatment modalities used. The maximum extent of resection should be safely attempted while minimizing the risk of surgically induced neurological injury.


Serum polychlorinated biphenyls, cytochrome P-450 1A1 polymorphisms, and risk of breast cancer in Connecticut women.
Recent epidemiologic studies have suggested that genetic polymorphisms in the cytochrome P-450 1A1 gene (CYP1A1) may affect the relation between environmental exposure to polychlorinated biphenyls (PCBs) and breast cancer risk. The authors report results from a case-control study evaluating the potential effect of gene-environment interaction between CYP1A1 and serum PCB levels on breast cancer risk among Caucasian women in Connecticut. The study included 374 case women with histologically confirmed breast cancer and 406 noncancerous controls with information on both serum PCB level and CYP1A1 genotype (1999-2002). Compared with women who had the homozygous wild-type CYP1A1 m2 genotype, significantly increased risks of breast cancer were found for women with the CYP1A1 m2 variant genotype (odds ratio (OR) = 2.1, 95% confidence interval (CI): 1.1, 3.9), especially postmenopausal women (OR = 2.4, 95% CI: 1.1, 5.0). Risks associated with the CYP1A1 m2 variant genotype were highest for all women (OR = 3.6, 95% CI: 1.5, 8.2) and postmenopausal women (OR = 4.3, 95% CI: 1.6, 12.0) with higher serum PCB levels (611-2,600 ng/g). The CYP1A1 m1 and m4 genotypes were not associated with breast cancer risk independently or in combination with PCB exposure. In summary, the CYP1A1 m2 genetic polymorphism was associated with increased risk of female breast cancer and may modify the relation between PCB exposure and breast cancer risk.


Genomic African and Native American Ancestry and Chagas Disease: The Bambui (Brazil) Epigen Cohort Study of Aging.
The influence of genetic ancestry on Trypanosoma cruzi infection and Chagas disease outcomes is unknown. We used 370,539 Single Nucleotide Polymorphisms (SNPs) to examine the association between individual proportions of African, European and Native American genomic ancestry with T. cruzi infection and related outcomes in 1,341 participants (aged ≥ 60 years) of the Bambui (Brazil) population-based cohort study of aging. Potential confounding variables included sociodemographic characteristics and an array of health measures. The prevalence of T. cruzi infection was 37.5% and 56.3% of those infected had a major ECG abnormality. Baseline T. cruzi infection was correlated with higher levels of African and Native American ancestry, which in turn were strongly associated with poor socioeconomic circumstances. Cardiomyopathy in infected persons was not significantly associated with African or Native American ancestry levels. Infected persons with a major ECG abnormality were at increased risk of 15-year mortality relative to their counterparts with no such abnormalities (adjusted hazard ratio = 1.80; 95% 1.41, 2.32). African and Native American ancestry levels had no significant effect modifying this association. Our findings indicate that African and Native American ancestry have no influence on the presence of major ECG abnormalities and had no influence on the ability of an ECG abnormality to predict mortality in older people infected with T. cruzi. In contrast, our results revealed a strong and independent association between prevalent T. cruzi infection and higher levels of African and Native American ancestry. Whether this association is a consequence of genetic background or differential exposure to infection remains to be determined.


Direct communication between osteocytes and acid-etched titanium implants with a sub-micron topography.
The osteocyte network, through the numerous dendritic processes of osteocytes, is responsible for sensing mechanical loading and orchestrates adaptive bone remodelling by communicating with both the osteoclasts and the osteoblasts. The osteocyte network in the vicinity of implant surfaces provides insight into the bone healing process around metallic implants. Here, we investigate whether osteocytes are able to make an intimate contact with topologically modified, but micrometre smooth (S a < 0.5 µm) implant surfaces, and if sub-micron topography alters the composition of the interfacial tissue. Screw shaped, commercially pure (cp-Ti) titanium implants with (i) machined (S a = ~0.2 µm), and (ii) two-step acid-etched (HF/HNO3 and H2SO4/HCl; S a = ~0.5 µm) surfaces were inserted in Sprague Dawley rat tibia and followed for 28 days. Both surfaces showed similar bone area, while the bone-implant contact was 73 % higher for the acid-etched surface. By resin cast etching, osteocytes were observed to maintain a direct intimate contact with the acid-etched surface. Although well mineralised, the interfacial tissue showed lower Ca/P and apatite-to-collagen ratios at the acid-etched surface, while mineral crystallinity and the carbonate-to-phosphate ratios were comparable for both implant surfaces. The interfacial tissue composition may therefore vary with changes in implant surface topography, independently of the amount of bone formed. Implant surfaces that influence bone to have higher amounts of organic matrix without affecting the crystallinity or the carbonate content of the mineral phase presumably result in a more resilient interfacial tissue, better able to resist crack development during functional loading than densely mineralised bone.


Utilitarianism and the pandemic.
There are no egalitarians in a pandemic. The scale of the challenge for health systems and public policy means that there is an ineluctable need to prioritize the needs of the many. It is impossible to treat all citizens equally, and a failure to carefully consider the consequences of actions could lead to massive preventable loss of life. In a pandemic there is a strong ethical need to consider how to do most good overall. Utilitarianism is an influential moral theory that states that the right action is the action that is expected to produce the greatest good. It offers clear operationalizable principles. In this paper we provide a summary of how utilitarianism could inform two challenging questions that have been important in the early phase of the pandemic: (a) Triage: which patients should receive access to a ventilator if there is overwhelming demand outstripping supply? (b) Lockdown: how should countries decide when to implement stringent social restrictions, balancing preventing deaths from COVID-19 with causing deaths and reductions in well-being from other causes? Our aim is not to argue that utilitarianism is the only relevant ethical theory, or in favour of a purely utilitarian approach. However, clearly considering which options will do the most good overall will help societies identify and consider the necessary cost of other values. Societies may choose either to embrace or not to embrace the utilitarian course, but with a clear understanding of the values involved and the price they are willing to pay.


Expression of cadherins and their undercoat proteins (alpha-, beta-, and gamma-catenins and p120) and accumulation of beta-catenin with no gene mutations in synovial sarcoma.
E-cadherin, the major intercellular adhesion molecule of epithelial cells, is important in determining the architecture of sarcomas, especially those showing epithelioid features. In addition to its role in cell adhesion, beta-catenin, a cadherin undercoat protein, has been shown to function as a downstream transcriptional activator of the Wnt/Wingless signaling pathway. In order to evaluate the significance of the cadherin cell adhesion system and the Wnt/Wingless signaling pathway in the morphogenesis and/or tumorigenesis of synovial sarcoma (a major type of sarcoma with epithelioid features), immunoreactivity for pan-cadherin, E-cadherin, and their undercoat proteins (alpha-, beta-,and gamma-catenins and p120) was evaluated in 15 synovial sarcomas. Immunoreactivity for pan-cadherin, E-cadherin, alpha-catenin, beta-catenin, and p120 was observed in all 15 specimens. Immunoreactivity for pan-cadherin was stronger than that for E-cadherin. Expression of gamma-catenin was detected in ten specimens. Although beta-catenin was observed only at the cell-cell boundaries in four specimens, it was present in the nucleus and cytoplasm and at the cell-cell boundaries in the other 11, suggesting constitutional activation of the Wnt/Wingless signaling pathway in synovial sarcoma. Direct sequencing for exon 3 of the beta-catenin gene, however, revealed no mutations in any of the 15 specimens. In conclusion, other types of cadherin besides E-cadherin, together with cadherin undercoat proteins, may play a role in cell adhesion in synovial sarcoma. Furthermore, mechanisms other than mutation of exon 3 of the beta-catenin gene may activate the Wnt/Wingless signaling pathway in this type of tumor.


Occlusal and oral health status of a group of 3-8-year-old South African black children.
This study determined the oral health status of a group of 3-8-year-old South African black children, comprising a total of 214 children from the townships of Garankuwa, Shosanguwe, Mabopane, Hebron and Erasmus who attended a school in Akasia, Greater Pretoria Metropolitan Substructure. The decayed, missing and filled teeth (dmft), oral hygiene status, dental IQ and malocclusion status were determined. The study found that the children's oral health status and occlusal status were unacceptable. The level of their dental IQ scores was low, their oral hygiene poor, and they were in urgent need of primary and secondary dental care. In addition they were in need of both preventive and interceptive orthodontic care. A national strategy to address primary dental health care is recommended.


Decrease in the CO2 uptake capacity in an ice-free Arctic Ocean basin.
It has been predicted that the Arctic Ocean will sequester much greater amounts of carbon dioxide (CO2) from the atmosphere as a result of sea ice melt and increasing primary productivity. However, this prediction was made on the basis of observations from either highly productive ocean margins or ice-covered basins before the recent major ice retreat. We report here a high-resolution survey of sea-surface CO2 concentration across the Canada Basin, showing a great increase relative to earlier observations. Rapid CO2 invasion from the atmosphere and low biological CO2 drawdown are the main causes for the higher CO2, which also acts as a barrier to further CO2 invasion. Contrary to the current view, we predict that the Arctic Ocean basin will not become a large atmospheric CO2 sink under ice-free conditions.


Characteristics of Chinese herbal medicine usage in ischemic heart disease patients among type 2 diabetes and their protection against hydrogen peroxide-mediated apoptosis in H9C2 cardiomyoblasts.
Evidence for long-term use of Chinese herbal medicine (CHM) as an adjuvant treatment in patients with type 2 diabetes (T2D) remains limited. This study aimed to assess the frequency of use, utilization patterns, and therapeutic effects of adjuvant CHM for ischemic heart disease (IHD) in patients with T2D in Taiwan. We identified 4620 IHD patients with T2D. After matching for age, gender, and insulin use, 988 subjects each were allocated to a CHM group and a non-CHM group. There were no differences in baseline characteristics except for comorbidities. The CHM group contained more cases with chronic obstructive pulmonary disease, hepatitis, ulcer disease, and hyperlipidemia. The cumulative survival probability was higher in CHM users than in matched non-CHM users aged 60 years or older (P < .0001, log rank test) regardless of gender (P = .0046 for men, P = .0010 for women, log rank test). Among the top 12 CHM combinations, Shu-Jing-Huo-Xue-Tang and Shao-Yao-Gan-Cao-Tang (13.6%) were the most common. This dual combination improved antiapoptotic activity in H2O2-exposed H9C2 cells by enhancing phosphorylation of glycogen synthase kinase-3β and p38 mitogen-activated protein kinase and could increase the survival of myocardial cells. Our study suggests that adjuvant CHM therapy may increase the survival probability and provides a comprehensive list for future investigations of the safety and efficacy of CHM for IHD patients with T2D.


Improved Geometry of Decellularized Tissue Engineered Heart Valves to Prevent Leaflet Retraction.
Recent studies on decellularized tissue engineered heart valves (DTEHVs) showed rapid host cell repopulation and increased valvular insufficiency developing over time, associated with leaflet shortening. A possible explanation for this result was found using computational simulations, which revealed radial leaflet compression in the original valvular geometry when subjected to physiological pressure conditions. Therefore, an improved geometry was suggested to enable radial leaflet extension to counteract for host cell mediated retraction. In this study, we propose a solution to impose this new geometry by using a constraining bioreactor insert during culture. Human cell based DTEHVs (n = 5) were produced as such, resulting in an enlarged coaptation area and profound belly curvature. Extracellular matrix was homogeneously distributed, with circumferential collagen alignment in the coaptation region and global tissue anisotropy. Based on in vitro functionality experiments, these DTEHVs showed competent hydrodynamic functionality under physiological pulmonary conditions and were fatigue resistant, with stable functionality up to 16 weeks in vivo simulation. Based on implemented mechanical data, our computational models revealed a considerable decrease in radial tissue compression with the obtained geometrical adjustments. Therefore, these improved DTEHV are expected to be less prone to host cell mediated leaflet retraction and will remain competent after implantation.


Factors associated with the occurrence of MRSA CC398 in herds of fattening pigs in Germany.
The purpose of this study was to investigate the prevalence of MRSA in herds of fattening pigs in different regions of Germany, and to determine factors associated with the occurrence of this pathogen. For this purpose pooled dust samples were collected, and a questionnaire covered information regarding herd characteristics and management practices. Samples were pre-enriched in high-salt medium followed by selective enrichment containing cefoxitin/aztreonam, and culturing. Presumptive colonies were confirmed by multiplex-PCR targeting nuc-, mecA- and 16S rRNA-genes. Isolates were spa- and SCCmec-, and in selected cases, multilocus sequence-typed. Susceptibilities to 13 antimicrobials were determined by broth microdilution. Statistical analysis was carried out using backward stepwise logistic regression to calculate odds ratios with the MRSA test result as the outcome and herd characteristics as categorical covariates. Overall, 152 of 290 (52%) fattening pig farms tested positive for MRSA. The prevalence in the east, north- and south-west of Germany ranged from 39 to 59%.t011 (66%) and t034 (23%) were the most commonly identified spa-types, and 85% of isolates carried SCCmec Type V. Identified spa-types were all associated with clonal complex CC398. Susceptibility testing revealed that all isolates were resistant to tetracycline. High resistance rates were also found for sulfamethoxazole/trimethoprim (40%), and quinupristin/dalfopristin (32%). In addition, 83% of strains displayed multidrug resistant (> 3 substance classes) phenotypes.Logistic regression revealed herd size (large farms OR: 5.4; CI: 2.7-11.2; p < 0.05), and production type (wean-to-finish OR: 4.0; CI: 1.6-10.4; p < 0.05) as risk factors associated with a positive MRSA finding in fattening pig operations. MRSA CC398 is widely distributed among herds of fattening pigs in Germany. Farm management plays a crucial role in the dissemination of MRSA with herd size, and production type representing potential major indicators.


Effect of luminal nucleotides on Cl- secretion and Na+ absorption in distal bronchi.
P2Y receptor agonists stimulate Cl- secretion across both normal and cystic fibrosis (CF) airway epithelia, and therefore have potential for use in the treatment of CF. Although CF pathology is manifest primarily in the distal airways, most studies of P2Y-receptor-mediated airway epithelial ion transport have used cells cultured from proximal regions. Here we report the results of studies of P2Y-receptor-mediated ion transport in distal bronchi isolated from porcine lungs, cannulated and perfused. A luminal microelectrode was used to record transepithelial potential difference (PD) and cable analysis was applied to determine resistance (Rt) and equivalent short-circuit current (I(SC)). Luminal UTP (100 micromol/l) transiently hyperpolarized PD (from -5.8+/-0.3 to -6.5+/-0.4 mV) and increased I(SC) (from 47+/-6 to 55+/-8 microA cm(-2)) before inhibiting PD to below the pre-UTP level (-5.0+/-0.4 mV). The decline was attenuated by pretreatment with amiloride, and additional treatment with bumetanide inhibited the initial hyperpolarization, suggesting that UTP stimulates Cl- secretion and inhibits Na+ absorption across distal bronchi. Luminal addition of P2Y1 [ADP, 2-methylthio-ATP (2MeSATP)] and P2Y6 (UDP) receptor agonists had no effect on ion transport. Pretreatment with thapsigargin (0.3 micromol/l) did not prevent the UTP-induced increase in PD and I(SC) but attenuated the secondary fall in PD. Pretreatment with BAPTA/AM (50 micromol/l), however, had no effect on the response to UTP. Additional studies of isolated bronchial epithelial cells using Fura-2 showed that UTP increases [Ca2+]in, and this increase is blocked by pretreatment with thapsigargin. These results suggest that in intact distal bronchi luminal UTP stimulates Cl- secretion by a Ca2+-independent mechanism and inhibits Na+ absorption by a Ca2+-dependent mechanism. Both effects are likely to favour increased hydration of the airway surface, and may therefore be beneficial in CF.


Are all sport activities equal? A systematic review of how youth psychosocial experiences vary across differing sport activities.
Models of sport development often support the assumption that young athletes' psychosocial experiences differ as a result of seemingly minor variations in how their sport activities are designed (eg, participating in team or individual sport; sampling many sports or specialising at an early age). This review was conducted to systematically search sport literature and explore how the design of sport activities relates to psychosocial outcomes. Systematic search, followed by data extraction and synthesis. The Preferred Reporting Items for Systematic reviews and Meta-Analyses guidelines were applied and a coding sheet was used to extract article information and code for risk of bias. Academic databases and manual search of peer-reviewed journals. Search criteria determined eligibility primarily based on the sample (eg, ages 7 through 17 years) and study design (eg, measured psychosocial constructs). 35 studies were located and were classified within three categories: (1) sport types, (2) sport settings, and (3) individual patterns of sport involvement. These studies represented a wide range of scores when assessed for risk of bias and involved an array of psychosocial constructs, with the most prevalent investigations predicting outcomes such as youth development, self-esteem and depression by comparing (1) team or individual sport participants and (2) youth with varying amounts of sport involvement. As variations in sport activities impact youth sport experiences, it is vital for researchers to carefully describe and study these factors, while practitioners may use the current findings when designing youth sport programmes.


An innovative technique for displaying three dimensional radiographic anatomy of synovial structures in the equine distal limb.
Radiography is the most commonly applied imaging modality in equine practice and forms an essential part of the diagnostic work-up of lame horses. Radiographic signs of musculoskeletal pathology are frequently localized at sites of soft tissue attachment, which are often not clearly visible on radiographs. Different lesions carry different prognoses and require a variety of treatments, and a good knowledge of the position of the synovial structures in the distal limb of the horse is essential for practitioners in the interpretation of radiographs. This study describes a new technique for creating three-dimensional (3D) models of the synovial structures and superimposing them onto radiographs for the purpose of teaching radiographic anatomy. A set of standard radiographs was acquired of the metacarpophalangeal and the distal interphalangeal joints of a fresh cadaver leg while the leg was positioned in a material-testing machine to mimic the weight-bearing horse. Computed tomography of the same regions was performed after injection of negative contrast medium into the joints. 3D reconstructions of the joints were created using grayscale thresholding and polynomial surface meshing in Mimics. The resulting 3D reconstructions were superimposed on top of the radiographs using Adobe Photoshop CS3 Extended, thus allowing the visualization of the joint anatomy in relation to the bone on all projections. The main advantage of this technique is that it allows synovial structures to be visualized on radiographs where they are normally indistinct, which will serve as a teaching aid for anatomy.


Single-cell variation leads to population invariance in NF-κB signaling dynamics.
The activation dynamics of nuclear factor (NF)-κB have been shown to affect downstream gene expression. On activation, NF-κB shuttles back and forth across the nuclear envelope. Many dynamic features of this shuttling have been characterized, and most features vary significantly with respect to ligand type and concentration. Here, we report an invariant feature with regard to NF-κB dynamics in cellular populations: the distribution--the average, as well as the variance--of the time between two nuclear entries (the period). We find that this period is conserved, regardless of concentration and across several different ligands. Intriguingly, the distributions observed at the population level are not observed in individual cells over 20-h time courses. Instead, the average period of NF-κB nuclear translocation varies considerably among single cells, and the variance is much smaller within a cell than that of the population. Finally, analysis of daughter-cell pairs and isogenic populations indicates that the dynamics of the NF-κB response is heritable but diverges over multiple divisions, on the time scale of weeks to months. These observations are contrary to the existing theory of NF-κB dynamics and suggest an additional level of control that regulates the overall distribution of translocation timing at the population level.


The effect of walking with an assistive device and using a wheelchair on school performance in students with myelomeningocele.
An alternating-condition, single-subject research design was used to examine the effect of ambulation on three measures of school performance in three students with myelomeningocele. The subjects, aged 9, 10, and 15 years, had a physiological cost index greater than 1.00 beats per meter when walking with crutches or a walker. Subjects propelled a wheelchair at school for 5 days, ambulated with crutches or a walker for 5 days, and propelled a wheelchair for an additional 5 days. Performance in reading fluency, visuomotor accuracy, and manual dexterity was assessed at the end of each school day. Results were graphed and analyzed using the two-standard-deviation band method. All subjects had significantly lower visuomotor accuracy scores during the assistive-device ambulation phase than during the wheelchair phases. Performance in manual dexterity during the assistive-device ambulation and wheelchair phases varied among the subjects. Reading fluency was not affected by method of mobility. The results suggest that the high energy cost of walking may have a negative effect on certain aspects of the subjects' school performance. [Franks CA, Palisano RJ, Darbee JC. The effect of walking with an assistive device and using a wheelchair on school performance in students with myelomeningocele.


IGFBP ratio confers resistance to IGF targeting and correlates with increased invasion and poor outcome in breast tumors.
To improve the significance of insulin-like growth factor-binding protein 5 (IGFBP-5) as a prognostic and potentially predictive marker in patients with breast cancer. Increased IGFBP-5 expression was identified in MCF-7 cells resistant (MCF-7R4) to the IGF-1R/insulin receptor (InsR) inhibitor BMS-536924 and its role examined by targeted knockdown and overexpression in multiple experimental models. Protein expression of IGFBP-5 was measured by immunohistochemistry in a cohort of 76 patients with breast cancer to examine correlative associations with invasive tumor fraction and outcome. The use of a combined IGFBP-5/IGFBP-4 (BPR) expression ratio was applied to predict anti-IGF-1R/InsR response in a panel of breast cancer lines and outcome in multiple breast tumor cohorts. IGFBP-5 knockdown decreased BMS-536924 resistance in MCF-7R4 cells, whereas IGFBP-5 overexpression in MCF-7 cells conferred resistance. When compared with pathologically normal reduction mammoplasty tissue, IGFBP-5 expression levels were upregulated in both invasive and histologically normal adjacent breast cancer tissue. In both univariate and multivariate modeling, metastasis-free survival, recurrence free survival (RFS), and overall survival (OS) were significantly associated with high IGFBP-5 expression. Prognostic power of IGFBP-5 was further increased with the addition of IGFBP-4 where tumors were ranked based upon IGFBP-5/IGFBP-4 expression ratio (BPR). Multiple breast cancer cohorts confirm that BPR (high vs. low) was a strong predictor of RFS and OS. IGFBP-5 expression is a marker of poor outcome in patients with breast cancer. An IGFBP-5/IGFBP-4 expression ratio may serve as a surrogate biomarker of IGF pathway activation and predict sensitivity to anti-IGF-1R targeting.


Effects of protein flexibility and active site water molecules on the prediction of sites of metabolism for cytochrome P450 2C19 substrates.
Structure-based prediction of sites of metabolism (SOMs) mediated by cytochrome P450s (CYPs) is of great interest in drug discovery and development. However, protein flexibility and active site water molecules remain a challenge for accurate SOM prediction. CYP2C19 is one of the major drug-metabolizing enzymes and has attracted considerable attention because of its polymorphism and capability of metabolizing ∼7% clinically used drugs. In this study, we systematically evaluated the effects of protein flexibility and active site water molecules on SOM prediction for CYP2C19 substrates. Multiple conformational sampling techniques including GOLD flexible residues sampling, molecular dynamics (MD) and tCONCOORD side-chain sampling were adopted for assessing the influence of protein flexibility on SOM prediction. The prediction accuracy could be significantly improved when protein flexibility was considered using the tCONCOORD sampling method, which indicated that the side-chain conformation was important for accurate prediction. However, the inclusion of the crystallographic or MD-derived water molecule(s) does not necessarily improve the prediction accuracy. Finally, a combination of docking results with SMARTCyp was found to be able to increase the SOM prediction accuracy.


SMAD proteins control DROSHA-mediated microRNA maturation.
MicroRNAs (miRNAs) are small non-coding RNAs that participate in the spatiotemporal regulation of messenger RNA and protein synthesis. Aberrant miRNA expression leads to developmental abnormalities and diseases, such as cardiovascular disorders and cancer; however, the stimuli and processes regulating miRNA biogenesis are largely unknown. The transforming growth factor beta (TGF-beta) and bone morphogenetic protein (BMP) family of growth factors orchestrates fundamental biological processes in development and in the homeostasis of adult tissues, including the vasculature. Here we show that induction of a contractile phenotype in human vascular smooth muscle cells by TGF-beta and BMPs is mediated by miR-21. miR-21 downregulates PDCD4 (programmed cell death 4), which in turn acts as a negative regulator of smooth muscle contractile genes. Surprisingly, TGF-beta and BMP signalling promotes a rapid increase in expression of mature miR-21 through a post-transcriptional step, promoting the processing of primary transcripts of miR-21 (pri-miR-21) into precursor miR-21 (pre-miR-21) by the DROSHA (also known as RNASEN) complex. TGF-beta- and BMP-specific SMAD signal transducers are recruited to pri-miR-21 in a complex with the RNA helicase p68 (also known as DDX5), a component of the DROSHA microprocessor complex. The shared cofactor SMAD4 is not required for this process. Thus, regulation of miRNA biogenesis by ligand-specific SMAD proteins is critical for control of the vascular smooth muscle cell phenotype and potentially for SMAD4-independent responses mediated by the TGF-beta and BMP signalling pathways.


Community based prevention programs targeting all injuries for children.
Community based models for injury prevention have become an accepted part of the overall injury control strategy. This systematic review of the scientific literature examines the evidence for their effectiveness in reducing all-cause injury in children 0-14 years of age. A comprehensive search of the literature was performed using the following study selection criteria: community based intervention study; children under 14 years; outcome measure was injury rates; and either a community control or an historical control was used in the design. Quality assessment and data abstraction were guided by a standardized procedure and performed independently by two authors. Data synthesis was in tabular and text form with meta-analysis not being possible due to the discrepancy in methods and measures between the studies. Thorough electronic and library search techniques yielded only nine formally evaluated community based all-cause child injury prevention programs that have reported actual injury outcomes. Of these nine studies, seven provided high level evidence where contemporary control communities were used for comparison; the remaining two used a pre and post-design or time trend analysis where historical data from the community were used as the comparison. Only three of the seven studies with contemporary control communities found significant effect of the intervention; the two studies without controls noted significant reductions in injury rates after the intervention period. There is a paucity of research from which evidence regarding the effectiveness of community based childhood injury prevention programs can be obtained and hence a clear need to increase the effort on developing this evidence base.


Development of a cell-based diabetic wound assay.
Chronic wounds require prolonged healthcare and adversely affect the quality of life of patients. They are particularly prominent in patients with diabetes, and their relative numbers are set to increase with the rise of diabetes within our population. Research is still needed to understand the factors leading to such wounds, to understand why they persist for such long periods of time, and also to develop new and efficacious treatment strategies. One problem facing this research is a lack of adequate animal models, as the current models do not truly reflect the human condition and often lead to much animal suffering. Hence, over the past four years, our group has been trying to develop a human-based in vitro diabetic wound model, which could be used as a high-throughput screening system to pre-screen potential chronic diabetic wound healing agents and to reduce unnecessary animal pain and suffering. To this end, we have isolated healthy and diseased skin fibroblasts from patient tissue biopsies. Crucially, to create a cell reporter system that can be widely used in the future, the cells were immortalised in order to escape senescence. By using microarray analysis, gene expression pattern differences have been identified between healthy and diseased cells, and disease-specific 'reporter' genes have been selected for further studies. The promoters of these reporter genes have been coupled to fluorescent reporter constructs and inserted back into the diseased fibroblasts, so that we now have proof-of-concept for a real-time diabetic reporter system for future exploitation.


Molecular epidemiological survey of Treponema pallidum in pregnant women in the Zhabei District of Shanghai.
To evaluate the epidemiological characteristics of syphilis, such as incidence, vertical transmission and genotypes, in pregnant women in the Zhabei District of Shanghai. In addition, the changes in genotypes and the efficiency of genotyping were evaluated. We screened 3022 pregnant women for syphilis in the Shanghai Zhabei Central Hospital. Whole blood, plasma, earlobe blood and specimens from genital ulcers or skin/mucosal lesions were collected from syphilis-positive patients. Samples were genotyped by analysing a combination of three genes: tpr, arp and tp0548 (the tpr/arp/tp0548 genotype). Clinical data were further collected to evaluate disease incidence, maternal-neonatal transmission and social factors. Out of 3022 pregnant women screened for syphilis, 41 were syphilis-positive. Of these, 43.9 % showed vertical transmission (18/41). The prevalence of syphilis in pregnant women was 1.32 %, higher than that in other districts in Shanghai in 2014. Genotyping was performed in 10/11 (90.9 %) samples of syphilis lesions, 8/41 (19.5 %) blood samples, 12/41 (29.3 %) plasma samples, and 20/39 (48.7 %) earlobe blood samples. The predominant genotype was 14d/f, followed by 15d/f, 13a/f, 13d/f and 9o/c. Genotype 13a/f was reported for the first time in the Shanghai area since 2010. The prevalence of syphilis in pregnant women in the Zhabei District was higher than that in other areas in Shanghai and the genotype is variable.


A Scoping Review of Constructs Measured Following Intervention for School Refusal: Are We Measuring Up?
Reviews of the effectiveness of interventions for school refusal (SR) rely upon well-conducted primary studies. Currently there are no guidelines for those conducting primary studies about the measurement of outcome following intervention for SR. Most people would agree that it is important to measure school attendance as an outcome but there has been little discussion about other constructs that warrant measurement. To facilitate this discussion and support the development of guidelines, we conducted a scoping review of constructs measured in studies evaluating intervention for SR. We screened the title and abstract of 3,213 publications found in peer-reviewed journals between 1980 and 2019. After full text review of 271 publications, 50 publications describing 51 studies were included. Results address the frequency with which constructs were measured, along with instruments used, informants, and time-points for measurement. Based on the results, we offer guidelines for choosing constructs to measure following intervention for SR and considerations for how to measure the constructs. Guidelines can increase consistency across primary studies, with benefits for future meta-analyses and international comparisons. They also provide support for practitioners contemplating routine evaluation of their interventions for SR. Ultimately, a core outcome set for SR can be developed.


Assessment of Family Caregiver Burden and Its Relationships Between Quality of Life, Arm Disability, Grip Strength, and Lymphedema Symptoms in Women with Postmastectomy Lymphedema: A Prospective Cross-Sectional Study.
The aim the study is to assess relationships between caregiver burden, quality of life, arm disability, grip strength, and lymphedema symptoms in women with postmastectomy lymphedema. Fifty-two patients with postmastectomy lymphedema were recruited. Burden Interview, Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire and European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC QLQ-30 and EORTC QLQ BRE-23) were used for the assessments of caregiver burden, arm disability, and quality of life. Lymphedema symptoms were questioned by Visual Analogue Scale (VAS) for pain, heaviness, and tension. Grip strength and arm volumes of the affected and unaffected sides were measured. Statistically significant positive correlations were detected between Burden Interview scores and DASH scores, VAS for pain and EORTC QLO-30 symptom scores. There were statistically negative correlations between Burden Interview and grip strength of affected side and unaffected side and lymphedema duration. Multiple linear regression was calculated to predict Burden Interview and EORTC QLO-30 functional and symptom scores based on age, DASH, stage, and duration of lymphedema. DASH score was significant predictors of Burden Interview while DASH score and Burden Interview scores were significant predictors of quality of life. Arm disability indicates caregiver burden and quality of life in patients with postmastectomy lymphedema while caregiver burden predicts the quality of life. These findings have implications for the management of lymphedema. We suggest that arm disability should be diagnosed and treated to improve caregiver burden and quality of life.


Three-dimensional comparison between the palatal forms in complete unilateral cleft lip and palate with and without Hotz plate from cheiloplasty to palatoplasty.
The palatal forms in 20 infants with a complete unilateral cleft lip, and palate (12 with a Hotz palate and 8 without, selected at random) were studied from birth until 18 months of age. Using techniques developed previously, the degree of curvature in the palate and the magnitude of migration of the maxillary segments were measured three-dimensionally. Furthermore, using a newly developed method to approximate a set of the points on the alveolar ridge to a circle in a plane, the form of the alveolar arch was evaluated. Results from the group with a Hotz plate revealed that the plate possesses four effects not seen in the group without a Hotz plate. The size of the palate was larger, and the sagittal gap between the two segments of the maxilla was smaller. These results suggest that the appliance could stimulate the growth of the segments and could prevent collapse of the maxillary arch from the force of lip closure. Third, the steepness of the segments toward the nasal cavity was smaller, possibly occurring because the appliance prevents tongue intrusion into the cleft. Fourth, the magnitude of migration of the lesser segment toward the cleft edge of the major segment was larger. This result suggests that the appliance could guide the growth of the maxillary segments to narrow the cleft width until 18 months of age.


Probiotics: Healthy bugs and nourishing elements of diet.
The use of probiotics is based on the concept that adding the right live microbes to the complex human system can result in physiological benefits. The benefit of fermented milk in human diet has been acknowledged since Vedic times; however, the scientific interest in this field was evoked by Ellie Metchinkoff who recommended that people should consume fermented milk containing lactobacilli to prolong their lives, as accelerated aging is due to autointoxication caused by the toxins produced by the gut microflora. They have been used to improve gastrointestinal health and their attractiveness has evinced interest to study their role in the promotion of oral health also. Studies have been widely carried out to establish the role of intestinal lactobacilli as probiotic to treat various gastrointestinal disorders, but only limited studies are available on the oral use of probiotics. The probiotic products usually contain lactobacilli and bifidobacteria, and their demand in the market is growing day by day. This paper provides an overview of various studies in the literature that emphasize on the role of probiotics to combat oral diseases and encourages more research in this field.


Bringing the commercial determinants of health out of the shadows: a review of how the commercial determinants are represented in conceptual frameworks.
The term 'commercial determinants of health' (CDOH) is increasingly focussing attention upon the role of tobacco, alcohol and food and beverage companies and others-as important drivers of non-communicable diseases (NCDs). However, the CDOH do not seem to be clearly represented in the most common social determinants of health (SDOH) frameworks. We review a wide range of existing frameworks of the determinants of health to determine whether and how commercial determinants are incorporated into current SDOH thinking. We searched for papers and non-academic reports published in English since 2000 describing influences on population health outcomes. We included documents with a formal conceptual framework or diagram, showing the integration of the different determinants. Forty-eight framework documents were identified. Only one explicitly included the CDOH in a conceptual diagram. Ten papers discussed the commercial determinants in some form in the text only and fourteen described negative impacts of commercial determinants in the text. Twelve discussed positive roles for the private sector in producing harmful commodities. Overall, descriptions of commercial determinants are frequently understated, not made explicit, or simply missing. The role of commercial actors as vectors of NCDs is largely absent or invisible in many of the most influential conceptual diagrams. Our current public health models may risk framing public health problems and solutions in ways that obscure the role that the private sector, in particular large transnational companies, play in shaping the broader environment and individual behaviours, and thus population health outcomes.


Perfusion seeding of channeled elastomeric scaffolds with myocytes and endothelial cells for cardiac tissue engineering.
The requirements for engineering clinically sized cardiac constructs include medium perfusion (to maintain cell viability throughout the construct volume) and the protection of cardiac myocytes from hydrodynamic shear. To reconcile these conflicting requirements, we proposed the use of porous elastomeric scaffolds with an array of channels providing conduits for medium perfusion, and sized to provide efficient transport of oxygen to the cells, by a combination of convective flow and molecular diffusion over short distances between the channels. In this study, we investigate the conditions for perfusion seeding of channeled constructs with myocytes and endothelial cells without the gel carrier we previously used to lock the cells within the scaffold pores. We first established the flow parameters for perfusion seeding of porous elastomer scaffolds using the C2C12 myoblast line, and determined that a linear perfusion velocity of 1.0 mm/s resulted in seeding efficiency of 87% +/- 26% within 2 hours. When applied to seeding of channeled scaffolds with neonatal rat cardiac myocytes, these conditions also resulted in high efficiency (77.2% +/- 23.7%) of cell seeding. Uniform spatial cell distributions were obtained when scaffolds were stacked on top of one another in perfusion cartridges, effectively closing off the channels during perfusion seeding. Perfusion seeding of single scaffolds resulted in preferential cell attachment at the channel surfaces, and was employed for seeding scaffolds with rat aortic endothelial cells. We thus propose that these techniques can be utilized to engineer thick and compact cardiac constructs with parallel channels lined with endothelial cells.


Heat shock protein 101 effects in A. thaliana: genetic variation, fitness and pleiotropy in controlled temperature conditions.
The Hsp100/ClpB heat shock protein family is ancient and required for high temperature survival, but natural variation in expression and its phenotypic effects is unexplored in plants. In controlled environment experiments, we examined the effects of variation in the Arabidopsis cytosolic AtHsp101 (hereafter Hsp101). Ten wild-collected ecotypes differed in Hsp101 expression responses across a 22 to 40 degrees C gradient. Genotypes from low latitudes expressed the least Hsp101. We tested fitness and pleiotropic consequences of varying Hsp101 expression in 'control' vs. mild thermal stress treatments (15/25 degrees C D/N vs. 15/25 degrees D/N plus 3 h at 35 degrees C 3 days/week). Comparing wild type and null mutants, wt Columbia (Col) produced approximately 33% more fruits compared to its Hsp101 homozygous null mutant. There was no difference between Landsberg erecta null mutant NIL (Ler) and wt Ler; wt Ler showed very low Hsp101 expression. In an assay of six genotypes, fecundity was a saturating function of Hsp101 content, in both experimental treatments. Thus, in addition to its essential role in acquired thermal tolerance, Hsp101 provides a substantial fitness benefit under normal growth conditions. Knocking out Hsp101 decreased fruit production, days to germination and days to bolting, total dry mass, and number of inflorescences; it increased transpiration rate and allocation to root mass. Root : total mass ratio decayed exponentially with Hsp101 content. This study shows that Hsp101 expression is evolvable in natural populations. Our results further suggest that Hsp101 is primarily an emergency high-temperature tolerance mechanism, since expression levels are lower in low-latitude populations from warmer climates. Hsp101 expression appears to carry an important trade-off in reduced root growth. This trade-off may select for suppressed expression under chronically high temperatures.


Older marijuana users in substance abuse treatment: Treatment settings for marijuana-only versus polysubstance use admissions.
Growing numbers of older adult marijuana users make understanding the marijuana-related treatment needs and treatment-related characteristics of this age group increasingly important. In this study, we examined four types of marijuana-involved admissions (marijuana as the only substance; marijuana as the primary substance with other secondary/tertiary substances; marijuana as the secondary substance; and marijuana as the tertiary substance) by treatment setting. Data came from the 2012-2017 Treatment Episode Data Set-Admissions (TEDS-A), which includes 851,652 admissions by those aged 55+. Using multinomial logistic regression analysis, we focused on the 120,286 marijuana-involved admissions to test the hypothesis that polysubstance use would be associated with a higher likelihood of using detoxification and rehabilitation settings than ambulatory/outpatient settings. Of all marijuana-involved admissions, 7.5% were marijuana-only, 12.7% were marijuana-primary, 58.4% were marijuana-secondary, and 21.4% were marijuana-tertiary admissions. Compared to marijuana-only admissions, admissions involving other substances were associated with a higher likelihood of detoxification and rehabilitation than ambulatory/outpatient treatment (e.g., RRR = 5.79, 95% CI = 5.08-6.61 for detoxification and RRR = 3.19, 95% CI = 2.89-3.52 for rehabilitation among marijuana-tertiary admissions). Referral source, first age of marijuana use, race/ethnicity, and homelessness were significant covariates. Given increasing numbers of older-adult marijuana users, healthcare providers should screen older adults for marijuana and other substance use, and substance abuse treatment programs should become more responsive to older adults' needs.


Symptomatic fracture incidence in southern Tasmania: does living in the country reduce your fracture risk?
There are limited data describing urban-rural differences in fracture incidence and the overall effect remains controversial. The aim of this study was to compare symptomatic fracture incidence occurring in geographically defined rural (n = 34619) and urban (n = 194974) populations of Southern Tasmania from July 1, 1997 to June 30, 1999. Fractures were ascertained by reviewing reports from all the radiology providers within the area. In the 2-year study time frame there were 3644 fractures in males and 2657 fractures in females. Fracture incidence was significantly higher in urban compared with rural populations in both sexes (male: RR 1.60, 95% CI 1.47-1.75; female: RR 1.77, 95% CI 1.58-1.98). This higher urban fracture incidence was present across all age groups and all fracture types with the exception of knee and pelvis fractures in males (although not all were statistically significant). In addition, urban men >50 years old had a higher fracture incidence than rural women >50 years old (RR 1.25, 95% CI 1.05-1.50), suggesting that in later life the factors responsible for the urban-rural difference are able to offset completely the effect of gender. While some of the reduced fracture incidence in the rural population may be explained by urban drift and underreporting of minor fractures such as foot fractures, the overall pattern of higher fracture risk was very consistent, suggesting a real difference in whole-of-life symptomatic fracture incidence. Further research at an individual level is required to determine what factors account for these large urban-rural differences, as they imply a substantial potential for fracture prevention.


Biomass Production in a Tallgrass Prairie Ecosystem Exposed to Ambient and Elevated CO"2.
Responses to elevated CO"2 have not been measured for natural grassland ecosystems. Global carbon budgets will likely be affected by changes in biomass production and allocation in the major terrestrial ecosystems. Whether ecosystems sequester or release excess carbon to the atmosphere will partly determine the extent and rate that atmospheric CO"2 concentration rises. Elevated CO"2 also may change plant community species composition and water status. We determined above- and belowground biomass production, plant community species composition, and measured and modeled water status of a tallgrass prairie ecosystem in Kansas exposed to ambient and twice-ambient CO"2 concentrations in open-top chambers during the entire growing season from 1989 through 1991. Dominant species were Andropogon gerardii, A. scoparius, and Sorghastrum nutans (C"4 metabolism) and Poa pratensis (C"3). Aboveground biomass and leaf area were estimated by periodic sampling throughout the growing season in 1989 and 1990. In 1991, peak biomass and leaf area were estimated by an early August harvest. Relative root production among treatments was estimated using root ingrowth bags which remained in place throughout the growing season. Latent heat flux was simulated with and without water stress. Botanical composition was estimated annually. Compared to ambient CO"2 levels, elevated CO"2 increased production of C"4 grass species, but not of C"3 grass species. composition of C"4 grasses did not change, but Poa pratensis (C"3) declined, and C"3 forbs increased in the stand with elevated CO"2 compared to ambient. Open-top chambers appeared to reduce latent heat flux and increase water-use efficiency similar to the elevated CO"2 treatment when water stress was not severe, but under severe water stress, the chamber effect on water-use efficiency was limited. In natural ecosystems with periodic moisture stress, increased water-use efficiency under elevated CO"2 apparently would have a greater impact on productivity irrespective of photosynthetic pathway.


A biodiversity approach in the neotropical erythrinidae fish, Hoplias malabaricus. Karyotypic survey, geographic distribution of cytotypes and cytotaxonomic considerations.
Hoplias malabaricus, a widely distributed neotropical freshwater fish, shows a conspicuous karyotypic diversification. An overview of this diversity is presented here comprising several Brazilian populations, and some others from Argentina, Uruguay and Surinam. Seven general cytotypes are clearly identified on the basis of their diploid number (2n = 39 to 2n = 42), chromosomal morphology and sex chromosome systems, which can be clustered into two major karyotypic groups. This clustering suggests that karyotype structure would be more informative than the diploid number regarding cytotype relationships in this fish group. While some cytotypes show a wide geographical distribution, some others appear to be endemic to specific hydrographic basins. Sympatric cytotypes can occur without detection of hybrid forms; this situation points to a lack of gene flow, a fact that is also reinforced by studies with genomic markers. The karyotypic data support the view that the nominal taxon H. malabaricus corresponds to a species complex comprising distinct evolutionary units, each with well-established chromosomal differences.


Cardiac transplantation in adults with congenital heart disease.
Due to increasing success with repair or palliation in childhood, there is a rapidly growing population of adult patients with complex congenital heart disease who may require transplantation. There remains little data on outcomes of cardiac transplantation in this group. 38 orthotopic cardiac transplants were performed in 37 patients (18 men) > or =18 years of age with congenital heart disease (CHD) from 1988 to 2009 in our institution. Outcomes were reviewed using medical records and transplant databases. 15 patients (41%) had univentricular and 22 (59%) biventricular physiology. The biggest group was transposition of the great arteries following atrial switch in eight patients (22%). Six (16%) had no previous surgical intervention. Mean age at transplant was 33.5 years (range 19.1-59.9 years). 11 patients (30%) required additional surgical procedures at transplant. 16 (43%) died, 12 early and 4 late deaths (1.8, 2.4, 2.7 and 7 years). Survival was 70% at 30 days, 68% at 1 year, 58% at 5 years and 53% at 10 and 15 years. Outcome improved in later eras with reduction in 30-day mortality from 50% to 18% and increase in 5-year survival from 50% to 69%. Two patients developed post-transplant lymphoproliferative disease. None required long-term renal replacement therapy. One patient was re-transplanted for cardiac allograft vasculopathy. While operative mortality following cardiac transplantation for adult congenital heart disease is higher than for other diagnostic groups, long-term survival is good and comparable to patients without CHD. Disappointing early results are improved with increasing experience.


Carbon disulfide. Just toxic or also bioregulatory and/or therapeutic?
The overview presented here has the goal of examining whether carbon disulfide (CS2) may play a role as an endogenously generated bioregulator and/or has therapeutic value. The neuro- and reproductive system toxicity of CS2 has been documented from its long-term use in the viscose rayon industry. CS2 is also used in the production of dithiocarbamates (DTCs), which are potent fungicides and pesticides, thus raising concern that CS2 may be an environmental toxin. However, DTCs also have recognized medicinal use in the treatment of heavy metal poisonings as well as having potency for reducing inflammation. Three known small molecule bioregulators (SMBs) nitric oxide, carbon monoxide, and hydrogen sulfide were initially viewed as environmental toxins. Yet each is now recognized as having intricate, though not fully elucidated, biological functions at concentration regimes far lower than the toxic doses. The literature also implies that the mammalian chemical biology of CS2 has broader implications from inflammatory states to the gut microbiome. On these bases, we suggest that the very nature of CS2 poisoning may be related to interrupting or overwhelming relevant regulatory or signaling process(es), much like other SMBs.


Depressive symptoms in older long-term colorectal cancer survivors: a population-based analysis using the SEER-Medicare healthcare outcomes survey.
Colorectal cancer survivorship has improved significantly over the last 20 years; however, few studies have evaluated depression among older colorectal cancer survivors, especially using a population-based sample. The aim of this study was to identify correlates for positive depression screen among colorectal cancer survivors who underwent potentially curative surgery. Using the 1998-2007 Surveillance, Epidemiology, and End-Result registry and the Medicare Health Outcome Survey linked dataset, we identified patients over 65 with pathology confirmed and resected colorectal cancer enrolled in Medicare. Using univariate and multiple variable analyses, we identified characteristics of patients with and without positive depression screen. Resected colorectal cancer patients (1785) (median age 77, 50.8 % female) were identified in the dataset with 278 (15.6 %) screening positive for symptoms of depression. Median time from diagnosis to survey was 62 months. On univariate analysis, larger tumor size, advanced cancer stage, and extent of resection were not correlates of depressive symptoms (all p > 0.05). After adjusting for confounders, income less than US$30,000 per year (OR 1.50, 1.02-2.22, 95 % CI, p = 0.042), non-white race (OR 1.51, 1.05-2.17, 95 % CI, p = 0.027), two or more comorbidities (OR 1.78, 1.25-2.52, 95 % CI, p = 0.001), and impairment in activities of daily living (OR 5.28, 3.67-7.60, 95 % CI, p < 0.001) were identified as independent correlates of depressive symptoms in colorectal cancer survivors. In the current study, socioeconomic status and features of physical health rather than tumor characteristics were associated with symptoms of depression among long-term colorectal cancer survivors.


Smooth muscle homeostasis in human atherosclerotic plaques through interleukin 15 signalling.
Interleukin (IL)-15 is a cytokine that has a broad tissue distribution and is important in maintaining homeostasis of cells and stability of tissues. When II-15 is also expressed by vascular smooth muscle cells (SMC), which are the dominant type of cells in most atherosclerotic plaques, it could be important in maintaining plaque tissue integrity and hence resistance of plaques towards development of clinically relevant complications such as plaque rupture and thrombosis. In this study, IL-15 and IL-15Rα in vitro expression by coronary artery SMC was investigated using RT -PCR and FACS analysis. Immunohistochemistry was used to study in situ expression of IL-15 and IL-15R by SMC of human carotid artery atherosclerotic plaques. Multiplex ligand-dependent probe amplification (MLPA) was used to investigate the mRNA expression of 40 pro- and anti inflammatory genes after stimulating coronary SMC with IL-15. We found that atherosclerotic SMC express both IL-15 and its receptor IL-15R, and TNF-γ and TNF-α enhance IL-15R expression in cultured SMC. MLPA studies on SMC revealed enhanced expression of PDGF beta mRNA after IL15 stimulation. In conclusion, our data suggest that IL-15 may contribute to atherosclerotic plaque integrity by stimulation of smooth muscle cells, probably in a PDGF dependent fashion.


Maternal exposure to Δ9-tetrahydrocannabinol impairs female offspring glucose homeostasis and endocrine pancreatic development in the rat.
Recent reports indicate that 7% of pregnant mothers in North America use cannabis. This is concerning given that in utero exposure to Δ9-tetrahydrocannabinol (Δ9-THC), the main psychoactive component in cannabis, causes fetal growth restriction and may alter replication and survival of pancreatic β-cells in the offspring. Accordingly, we hypothesized that maternal exposure to Δ9-THC during pregnancy would impair postnatal glucometabolic health of offspring. To test this hypothesis, pregnant Wistar rats were treated with daily intraperitoneal injections of either 3 mg/kg Δ9-THC or vehicle from gestational day 6 to birth. Offspring were subsequently challenged with glucose and insulin at 5 months of age to assess glucose tolerance and peripheral muscle insulin sensitivity. Female offspring exposed to Δ9-THC in utero were glucose intolerant, associated with blunted insulin response in muscle and increased serum insulin concentration 15 min after glucose challenge. Additionally, pancreata from male and female offspring were harvested at postnatal day 21 and 5 months of age for assessment of endocrine pancreas morphometry by immunostaining. This analysis revealed that gestational exposure to Δ9-THC reduced the density of islets in female, but not male, offspring at postnatal day 21 and 5 months, culminating in reduced β-cell mass at 5 months. These results demonstrate that fetal exposure to Δ9-THC causes female-specific impairments in glucose homeostasis, raising concern regarding the metabolic health of offspring, particularly females, exposed to cannabis in utero.


Molecular characterization and ligand binding specificity of the PDZ domain-containing protein GIPC3 from Schistosoma japonicum.
Schistosomiasis is a serious global health problem that afflicts more than 230 million people in 77 countries. Long-term mass treatments with the only available drug, praziquantel, have caused growing concerns about drug resistance. PSD-95/Dlg/ZO-1 (PDZ) domain-containing proteins are recognized as potential targets for the next generation of drug development. However, the PDZ domain-containing protein family in parasites has largely been unexplored. We present the molecular characteristics of a PDZ domain-containing protein, GIPC3, from Schistosoma japonicum (SjGIPC3) according to bioinformatics analysis and experimental approaches. The ligand binding specificity of the PDZ domain of SjGIPC3 was confirmed by screening an arbitrary peptide library in yeast two-hybrid (Y2H) assays. The native ligand candidates were predicted by Tailfit software based on the C-terminal binding specificity, and further validated by Y2H assays. SjGIPC3 is a single PDZ domain-containing protein comprised of 328 amino acid residues. Structural prediction revealed that a conserved PDZ domain was presented in the middle region of the protein. Phylogenetic analysis revealed that SjGIPC3 and other trematode orthologues clustered into a well-defined cluster but were distinguishable from those of other phyla. Transcriptional analysis by quantitative RT-PCR revealed that the SjGIPC3 gene was relatively highly expressed in the stages within the host, especially in male adult worms. By using Y2H assays to screen an arbitrary peptide library, we confirmed the C-terminal binding specificity of the SjGIPC3-PDZ domain, which could be deduced as a consensus sequence, -[SDEC]-[STIL]-[HSNQDE]-[VIL]*. Furthermore, six proteins were predicted to be native ligand candidates of SjGIPC3 based on the C-terminal binding properties and other biological information; four of these were confirmed to be potential ligands using the Y2H system. In this study, we first characterized a PDZ domain-containing protein GIPC3 in S. japonicum. The SjGIPC3-PDZ domain is able to bind both type I and II ligand C-terminal motifs. The identification of native ligand will help reveal the potential biological function of SjGIPC3. These data will facilitate the identification of novel drug targets against S. japonicum infections.


Associations between depressive symptoms, self-efficacy, eating styles, exercise and body mass index in women.
This article explores cross-sectional associations between depressive symptoms and body mass index (BMI) in women working in schools in the Greater New Orleans area. Self-efficacy for eating and exercise, eating styles, and exercise are examined as potential pathways. This is a secondary data analysis of 743 women who were participating in a workplace wellness randomized controlled trial to address environmental factors influencing eating and exercise behaviors using baseline data prior to the intervention. BMI was the primary outcome examined. Path analysis suggested that increased depressive symptoms were associated with increased BMI in women. Indirect effects of depressive symptoms on BMI were found for increased healthy eating self-efficacy, increased emotional eating, and decreased exercise self-efficacy. The association between greater healthy eating self efficacy and BMI was unexpected, and may indicate a suppressor effect of eating self-efficacy in the relationship between depressive symptoms and BMI in women. The findings suggest the importance of depressive symptoms to BMI in women. Targets for interventions to reduce BMI include targeting depressive symptoms and related sequelae including self-efficacy for exercise, and emotional eating. Further investigation of eating self-efficacy and BMI are recommended with particular attention to both efficacy for health eating and avoidance of unhealthy foods.


Residential Mobility, Family Structure, and Completion of Upper Secondary Education - A Registry-Based Cohort Study of the Norwegian Adolescent Population.
Longitudinal studies exploring the complex interplay between family structures and residential mobility on educational achievement and failure are lacking. We investigate the interplay between the number of residential moves during late childhood, parental education level, family living situation, and the probability of completing upper secondary education. Detailed longitudinal data for a random sample of 30% of the entire Norwegian population born 1982 to 1989 (N = 121,247) and information on all their relocations between Norwegian enumeration districts from ages 10 to 18 years were extracted from the Norwegian population registries. Family structures were grouped into four intersectional family strata defined by combining categories of parental education level (distinguishing poorly educated and well-educated families) and the family's living situation (comparing non-intact families with intact families). We applied two-level logistic regression models, which incorporated individual and family contextual factors, to estimate possible differences in completion rates of upper secondary education. Non-completion of secondary education (which constitutes 29% of the study sample) increases incrementally with the number of residential changes across all four family structures, but this effect was not distributed evenly between the different family strata. Individuals in "well-educated, intact families" seem to be least affected by residential moves. On the other hand, the highest disadvantage of frequent moves was among adolescents in the stratum "poorly educated, intact families." In poorly educated families the probabilities of completing secondary school among non-intact and intact families converge toward each other as the number of moves increase. About 43% of the variation in school completion may be attributed to differences between families. The highest risk of school non-completion was found among adolescents in poorly educated families, which accounted for 74% of the non-completers. We demonstrated underlying links between residential mobility and family structures on non-completion of upper secondary education. The adverse effect of frequent moves calls for attention in schools, public health agencies, and housing policies. The findings should be considered in a life course perspective, as the accumulation of unfavorable conditions during childhood and adolescence tends to constrict future prospects in terms of health and quality of life.


Inferring introgression using RADseq and DFOIL : Power and pitfalls revealed in a case study of spiny lizards (Sceloporus).
Introgression is now commonly reported in studies across the Tree of Life, aided by recent advancements in data collection and analysis. Nevertheless, researchers working with nonmodel species lacking reference genomes may be stymied by a mismatch between available resources and methodological demands. In this study, we demonstrate a fast and simple approach for inferring introgression using RADseq data, and apply it to a case study involving spiny lizards (Sceloporus) from northeastern México. First, we find evidence for recurrent mtDNA introgression between the two focal species based on patterns of mito-nuclear discordance. We then test for nuclear introgression by exhaustively applying the "five-taxon" D-statistic (DFOIL ) to all relevant individuals sampled for RADseq data. In our case, this exhaustive approach (dubbed "ExDFOIL ") entails testing up to ~250,000 unique four-taxon combinations of individuals across species. To facilitate use of this ExDFOIL approach, we provide scripts for many relevant tasks, including the selection of appropriate four-taxon combinations, execution of DFOIL tests in parallel and visualization of introgression results in phylogenetic and geographic space. Using ExDFOIL , we find evidence for ancient introgression between the focal species. Furthermore, we reveal geographic variation in patterns of introgression that is consistent with patterns of mito-nuclear discordance and with recurrent introgression. Overall, our study demonstrates that the combination of DFOIL and RADseq data can effectively detect introgression under a variety of sampling conditions (for individuals, populations and loci). Importantly, we also find evidence that batch-specific error and linkage in RADseq data may mislead inferences of introgression under certain conditions.


Endoscopic endonasal surgery for epidermoid and dermoid cysts: a 10-year experience.
OBJECTIVEEpidermoid and dermoid cysts may be found along the cranial base and are commonly resected via open transcranial approaches. The use of endoscopic endonasal approaches for resection of these tumors has been rarely reported.METHODSThe authors retrospectively reviewed the medical records of 21 patients who underwent endoscopic endonasal surgery for epidermoid and dermoid cyst resection at the University of Pittsburgh Medical Center between January 2005 and June 2014. Surgical outcomes and variables that might affect the extent of resection and complications were analyzed.RESULTSTotal resection (total removal of cyst contents and capsule) was achieved in 8 patients (38.1%), near-total resection (total removal of cyst contents, incomplete removal of cyst capsule) in 9 patients (42.9%), and subtotal resection (incomplete removal of cyst contents and capsule) in 4 patients (19%). Larger cyst volume (≥ 3 cm3) and intradural location (15 cysts) were significantly associated with nontotal resection (p = 0.008 and 0.0005, respectively). In the whole series, surgical complications were seen in 6 patients (28.6%). No complications were observed in patients with extradural cysts. Among the 15 patients with intradural cysts, the most common surgical complication was postoperative CSF leak (5 patients, 33.3%), followed by postoperative intracranial infection (4 patients, 26.7%). Larger cysts and postoperative CSF leak were associated with intracranial infection (p = 0.012 and 0.028, respectively). Subtotal resection was marginally associated with intracranial infection when compared with total resection (p = 0.091). All patients with neurological symptoms improved postoperatively with the exception of 1 patient with unchanged abducens nerve palsy.CONCLUSIONSEndoscopic endonasal approaches may be effectively used for resection of epidermoid and dermoid cysts in carefully selected cases. These approaches are recommended for cases in which a total or near-total resection is possible in addition to a multilayer cranial base reconstruction with vascularized tissue to minimize the risk of intracranial infection.


Building community capacity using web-supported work-based learning.
Health visitors are a central component of policy to create strong, stable families and communities. The programme which is described here is intended to facilitate existing health visitors to gain confidence and extend or renew their skills in building community capacity (BCC). Networking and relationships are essential to effective community development. These are key skills for the health visitor, which along with professional principles support community capacity building. Learning in this programme is self-directed, supported by web-based resources over a 24 week period. Learning mainly takes place in practice. It involves carrying out a work based project through to completion. Participants register online, and follow a series of six phases. Evaluation of the pilot took place during 2011. Three main areas for improvement were identified: reflective software; signposting access to resources; and dealing with workload pressures. Community engagement for health improvement remains an important element of the vision for health visiting. The programme described is a core resource through which health visitors can build the skills and confidence of community groups and staff in other agencies to make a difference to health and wellbeing.


Optimization of the matrix inversion tomosynthesis (MITS) impulse response and modulation transfer function characteristics for chest imaging.
Matrix inversion tomosynthesis (MITS) uses linear systems theory, along with a priori knowledge of the imaging geometry, to deterministically distinguish between true structure and overlying tomographic blur in a set of conventional tomosynthesis planes. In this paper we examine the effect of total scan angle (ANG), number of input projections (N), and plane separation/number of reconstructed planes (NP) on the MITS impulse response (IR) and modulation transfer function (MTF), with the purpose of optimizing MITS imaging of the chest. MITS IR and MTF data were generated by simulating the imaging of a very thin wire, using various combinations of ANG, N, and NP. Actual tomosynthesis data of an anthropomorphic chest phantom were acquired with a prototype experimental system, using the same imaging parameter combinations as those in the simulations. Thoracic projection data from two human subjects were collected for corroboration of the system response analysis in vivo. Results suggest that ANG=20 degrees, N=71, NP=69 is the optimal combination for MITS chest imaging given the inherent constraints of our prototype system. MITS chest data from human subjects demonstrates that the selected imaging strategy can effectively produce high-quality MITS thoracic images in vivo.


Ultrafast dynamics of myoglobin without the distal histidine: stimulated vibrational echo experiments and molecular dynamics simulations.
Ultrafast protein dynamics of the CO adduct of a myoglobin mutant with the polar distal histidine replaced by a nonpolar valine (H64V) have been investigated by spectrally resolved infrared stimulated vibrational echo experiments and molecular dynamics (MD) simulations. In aqueous solution at room temperature, the vibrational dephasing rate of CO in the mutant is reduced by approximately 50% relative to the native protein. This finding confirms that the dephasing of the CO vibration in the native protein is sensitive to the interaction between the ligand and the distal histidine. The stimulated vibrational echo observable is calculated from MD simulations of H64V within a model in which vibrational dephasing is driven by electrostatic forces. In agreement with experiment, calculated vibrational echoes show slower dephasing for the mutant than for the native protein. However, vibrational echoes calculated for H64V do not show the quantitative agreement with measurements demonstrated previously for the native protein.


Forerunner genes contiguous to RB1 contribute to the development of in situ neoplasia.
We used human bladder cancer as a model system and the whole-organ histologic and genetic mapping strategy to identify clonal genetic hits associated with growth advantage, tracking the evolution of bladder cancer from intraurothelial precursor lesions. Six putative chromosomal regions critical for clonal expansion of intraurothelial neoplasia and development of bladder cancer were identified by using this approach. Focusing on one of the regions, which includes the model tumor suppressor RB1, we performed allelotyping of single-nucleotide polymorphic sites and identified a 1.34-Mb segment around RB1 characterized by a loss of polymorphism associated with the initial expansion of in situ neoplasia. This segment contains several positional candidate genes referred to by us as forerunner genes that may contribute to such expansion. We subsequently concentrated our efforts on the two neighbor genes flanking RB1, namely ITM2B and CHC1L, as well as P2RY5, which is located inside RB1. Here, we report that ITM2B and P2RY5 modulated cell survival and were silenced by methylation or point mutations, respectively, and thus by functional loss may contribute to the growth advantage of neoplasia. We also show that homozygous inactivation of P2RY5 was antecedent to the loss of RB1 during tumor development, and that nucleotide substitutions in P2RY5 represent a cancer predisposing factor.


Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone.
We constructed an infectious molecular clone of acquired immunodeficiency syndrome-associated retrovirus. Upon transfection, this clone directed the production of infectious virus particles in a wide variety of cells in addition to human T4 cells. The progeny, infectious virions, were synthesized in mouse, mink, monkey, and several human non-T cell lines, indicating the absence of any intracellular obstacle to viral RNA or protein production or assembly. During the course of these studies, a human colon carcinoma cell line, exquisitely sensitive to DNA transfection, was identified.


Soluble lipopolysaccharide receptor (CD14) is released via two different mechanisms from human monocytes and CD14 transfectants.
The receptor for lipopolysaccharide LPS (CD14) exists in a membrane-associated (mCD14) and a soluble form (sCD14). Previous studies indicate that monocytes produce sCD14 by limited proteolysis of the membrane-bound receptor. In this study we demonstrate that human monocytes also produce sCD14 by a protease-independent mechanism. To investigate the molecular nature of this second pathway we studied sCD14 formation in the monocytic cell line Mono Mac 6 (MM6) and in CD14 transfectants. Both MM6 and the CD14 transfectants constitutively produce sCD14 by a protease-independent mechanism. Structural analysis of sCD14 produced by the CD14 transfectants reconfirmed the presence of the COOH terminus predicted from the cDNA. Since glycosylphosphatidylinositol anchor attachment is associated with the removal of a hydrophobic C-terminal signal peptide, our finding demonstrates that the transfectants secrete sCD14 which escaped this posttranslational modification. Identical results obtained for sCD14 derived from peritoneal dialysis fluid of a patient with kidney dysfunction show the in vivo relevance of this pathway for sCD14 production.


Alterations in the Geometry, Fiber Orientation, and Mechanical Behavior of the Lumbar Intervertebral Disc by Nucleus Swelling.
Soft tissues observed in clinical medical images are often prestrained in tension by internal pressure or tissue hydration. For a native disc, nucleus swelling occurs in equilibrium with osmotic pressure induced by the high concentration of proteoglycan in the nucleus. The objective of this computational study was to investigate the effects of nucleus swelling on disc geometry, fiber orientation, and mechanical behavior by comparing those of prestrained and zero-pressure (unswelled) discs. Thermoelastic analysis techniques were repurposed in order to determine the zero-pressure disc geometry which, when pressurized, matches the prestrained disc geometry observed in clinical images. The zero-pressure geometry was then used in simulations to approximately represent a degenerated disc, which loses the ability of nucleus swelling but has not undergone distinct soft tissue remodeling/disruption. Our simulation results demonstrated that the loss of nucleus swelling caused a slight change in the disc geometry and fiber orientation, but a distinct deterioration in the resistance to intervertebral rotations including sagittal bending, lateral bending, and axial torsion. Different from rotational loading, in compression (with a displacement of 0.45 mm applied), a much larger stiffness (3.02 KN/mm) and a greater intradiscal pressure (IDP) (0.61 MPa) were measured in the zero-pressure disc, compared to the prestrained disc (1.41 KN/mm and 0.52 MPa). This computational study could be useful to understand mechanisms of disc degeneration, and guide the future design of disc tissue engineering material and biomimic disc implants.